LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment

Abstract Background Inflammation often causes irreversible damage to dental pulp tissue. Dental pulp stem cells (DPSCs), which have multidirectional differentiation ability, play critical roles in the repair and regeneration of pulp tissue. However, the presence of proinflammatory factors can affect...

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Main Authors: Yunfeng Ma, Xinyuan Liu, Ruoxi Dai, Quanli Li, Chris Ying Cao
Format: Article
Language:English
Published: BMC 2024-12-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-024-04075-7
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author Yunfeng Ma
Xinyuan Liu
Ruoxi Dai
Quanli Li
Chris Ying Cao
author_facet Yunfeng Ma
Xinyuan Liu
Ruoxi Dai
Quanli Li
Chris Ying Cao
author_sort Yunfeng Ma
collection DOAJ
description Abstract Background Inflammation often causes irreversible damage to dental pulp tissue. Dental pulp stem cells (DPSCs), which have multidirectional differentiation ability, play critical roles in the repair and regeneration of pulp tissue. However, the presence of proinflammatory factors can affect DPSCs proliferation, differentiation, migration, and other functions. LL-37 is a natural cationic polypeptide that inhibits lipopolysaccharide (LPS) activity, enhances cytokine production, and promotes the migration of stem cells. However, the potential of LL-37 in regenerative endodontics remains unknown. This study aimed to investigate the regulatory role of LL-37 in promoting the migration and odontogenic differentiation of DPSCs within an inflammatory microenvironment. These findings establish an experimental foundation for the regenerative treatment of pulpitis and provide a scientific basis for its clinical application. Materials and methods DPSCs were isolated via enzyme digestion combined with the tissue block adhesion method and identified via flow cytometry. The impact of LL-37 on the proliferation of DPSCs was evaluated via a CCK-8 assay. The recruitment of DPSCs was assessed through a transwell assay. The mRNA expression levels of inflammatory and aging-related genes were assessed via reverse transcription‒polymerase chain reaction (RT‒PCR), western blotting, and enzyme‒linked immunosorbent assay (ELISA). The odontogenic differentiation of DPSCs was assessed through alkaline phosphatase (ALP) staining, alizarin red staining, and RT‒PCR analysis. Results LL-37 has the potential to enhance the migration of DPSCs. In an inflammatory microenvironment, LL-37 can suppress the expression of genes associated with inflammation and aging, such as TNF-α, IL-1β, IL-6, P21, P38 and P53. Moreover, it promotes odontogenic differentiation in DPSCs by increasing ALP activity, increasing calcium nodule formation, and increasing the expression of dentin-related genes such as DMP1, DSPP and BSP. Conclusion These findings suggest that the polypeptide LL-37 facilitates the migration of DPSCs and plays a crucial role in resolving inflammation and promoting cell differentiation within an inflammatory microenvironment. Consequently, LL-37 has promising potential as an innovative therapeutic approach for managing inflammatory dental pulp conditions.
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spelling doaj-art-647ba75ce0c24afea41d249cdf854a502025-08-20T01:57:11ZengBMCStem Cell Research & Therapy1757-65122024-12-0115111410.1186/s13287-024-04075-7LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironmentYunfeng Ma0Xinyuan Liu1Ruoxi Dai2Quanli Li3Chris Ying Cao4Key Lab. of Oral Diseases Research, College and Hospital of Stomatology, Anhui Medical UniversityKey Lab. of Oral Diseases Research, College and Hospital of Stomatology, Anhui Medical UniversityDepartment of Comprehensive Care, Tufts University School of Dental MedicineDepartment of Stomatology, Longgang Otorhinolaryngology Hospital of Shenzhen, Institute of Oral ScienceKey Lab. of Oral Diseases Research, College and Hospital of Stomatology, Anhui Medical UniversityAbstract Background Inflammation often causes irreversible damage to dental pulp tissue. Dental pulp stem cells (DPSCs), which have multidirectional differentiation ability, play critical roles in the repair and regeneration of pulp tissue. However, the presence of proinflammatory factors can affect DPSCs proliferation, differentiation, migration, and other functions. LL-37 is a natural cationic polypeptide that inhibits lipopolysaccharide (LPS) activity, enhances cytokine production, and promotes the migration of stem cells. However, the potential of LL-37 in regenerative endodontics remains unknown. This study aimed to investigate the regulatory role of LL-37 in promoting the migration and odontogenic differentiation of DPSCs within an inflammatory microenvironment. These findings establish an experimental foundation for the regenerative treatment of pulpitis and provide a scientific basis for its clinical application. Materials and methods DPSCs were isolated via enzyme digestion combined with the tissue block adhesion method and identified via flow cytometry. The impact of LL-37 on the proliferation of DPSCs was evaluated via a CCK-8 assay. The recruitment of DPSCs was assessed through a transwell assay. The mRNA expression levels of inflammatory and aging-related genes were assessed via reverse transcription‒polymerase chain reaction (RT‒PCR), western blotting, and enzyme‒linked immunosorbent assay (ELISA). The odontogenic differentiation of DPSCs was assessed through alkaline phosphatase (ALP) staining, alizarin red staining, and RT‒PCR analysis. Results LL-37 has the potential to enhance the migration of DPSCs. In an inflammatory microenvironment, LL-37 can suppress the expression of genes associated with inflammation and aging, such as TNF-α, IL-1β, IL-6, P21, P38 and P53. Moreover, it promotes odontogenic differentiation in DPSCs by increasing ALP activity, increasing calcium nodule formation, and increasing the expression of dentin-related genes such as DMP1, DSPP and BSP. Conclusion These findings suggest that the polypeptide LL-37 facilitates the migration of DPSCs and plays a crucial role in resolving inflammation and promoting cell differentiation within an inflammatory microenvironment. Consequently, LL-37 has promising potential as an innovative therapeutic approach for managing inflammatory dental pulp conditions.https://doi.org/10.1186/s13287-024-04075-7Dental pulp stem cellsPulpitisPolypeptideLipopolysaccharideOdontogenic differentiation
spellingShingle Yunfeng Ma
Xinyuan Liu
Ruoxi Dai
Quanli Li
Chris Ying Cao
LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
Stem Cell Research & Therapy
Dental pulp stem cells
Pulpitis
Polypeptide
Lipopolysaccharide
Odontogenic differentiation
title LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
title_full LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
title_fullStr LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
title_full_unstemmed LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
title_short LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
title_sort ll 37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment
topic Dental pulp stem cells
Pulpitis
Polypeptide
Lipopolysaccharide
Odontogenic differentiation
url https://doi.org/10.1186/s13287-024-04075-7
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